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Immunotherapy represents the future of clinical cancer treatment. Chimeric antigen receptor- macrophages (CAR-MA) are able to recognize and attack target cells, indicating macrophages'activation and engulfment is a promising therapeutic avenue. The establishment of an in vitro phagocytosis assay provides a fast and reliable method to prescreen and assess the phagocytic capacity of CAR-MA. Eating assay could be used to screen successful anti-cancer drugs in vitro and reduce the time and cost expended on animal models. Creative Biolabs provides quantitative methods to accurately measure phagocytosis.

Introduction of Macrophage Phagocytosis

Macrophage phagocytosis can be triggered by diverse receptor-ligand interactions to clear pathogens and dead cells from a host. When a macrophage ingests a pathogen, the pathogen becomes trapped in a phagosome, which then fuses with a lysosome. Phagocytosis can be divided into four main steps:

Recognition of the target particle
Signaling to activate the internalization machiner
Phagosome formation
Phagolysosome maturation

Fig.1 Targeted phagocytosis of cancer cells by CAR-MAs and activation of T cells. (Chen, 2021)

Approaches

The idea of fitting macrophages with CAR constructs confers target antigen specificity and promotes engineered macrophages to efficaciously target solid tumors. Therapeutic activation of macrophage phagocytosis can restrain tumor growth through phagocytic clearance of tumor cells and activation of the adaptive immune response. Therefore, there is a growing demand for methods of measuring cancer cell phagocytosis. Many ways of assaying phagocytosis exist that utilize a variety of phagocytic targets with different combinations of receptor-ligand interactions.

Creative Biolabs provides two phagocytosis assays, a fluorescent microscopic assay, and a flow cytometric assay. These phagocytosis assays are suitable for multiple types of tumors or phagocytes and can be used to evaluate the efficacy of various phagocytosis enhancers.

pHrodo-SE dye is utilized to label cancer cells and visualize engulfed cancer cells when at the low pH of the phagolysosome. This technique allows for the straightforward quantification of phagocytosis of cancer cells using fluorescence microscopy.
We analyze the fluorescent signal of labeling tumor cells co-cultured with macrophages by flow cytometry. Flow cytometry enables a rapid, quantitative, and accurate measurement of phagocytosis efficiency.

Fig.2 The schematic methods of cancer phagocytosis assay. (Creative Biolabs)

A better understanding of the interactions between tumor cells and phagocytes through in vitro phagocytosis assays may contribute to the identification of novel targets for cancer immunotherapy. Creative Biolabs provides a comprehensive service to detect CAR-MA phagocytosis and its efficiency. If you are interested in our services, please feel free to contact us.

References

Chen, Y.; et al. CAR-macrophage: a new immunotherapy candidate against solid tumors. Biomed Pharmacother. 2021, 139: 111605.
Nam, G.H.; et al. An optimized protocol to determine the engulfment of cancer cells by phagocytes using flow cytometry and fluorescence microscopy. J Immunol Methods. 2019, 470: 27-32.